An improved green fluorescent protein gene as a vital marker in plants.

Abstract:

:A synthetic green fluorescent protein (GFP) gene (pgfp) was constructed to improve GFP expression in plants. Corn and tobacco protoplast transient assays showed that pgfp gave about 20-fold brighter fluorescence than the wild-type gene (gfp). Replacement of the serine at position 65 with a threonine (S65Tpgfp) or a cysteine (S65Cpgfp) yielded 100- to 120-fold brighter fluorescence than wild-type gfp upon excitation with 490-nm light. Incorporation of a plant intron into the coding region yielded an additional 1.4-fold improvement, for a cumulative improvement of about 150-fold in fluorescence at 490-nm excitation. Various versions of pgfp were also stably introduced into corn, wheat, tobacco, and Arabidopsis plants. Bright-green fluorescence was observed with a fluorescence microscope in virtually all examined tissues of transgenic monocots and dicots. In the case of Arabidopsis, expression of the pgfp gene under the enhanced 355 promoter of the cauliflower mosaic virus produced green fluorescence that was readily detectable by eye using a hand-held, long-wave ultraviolet lamp and/or a black-light source.

journal_name

Plant Physiol

journal_title

Plant physiology

authors

Pang SZ,DeBoer DL,Wan Y,Ye G,Layton JG,Neher MK,Armstrong CL,Fry JE,Hinchee MA,Fromm ME

doi

10.1104/pp.112.3.893

subject

Has Abstract

pub_date

1996-11-01 00:00:00

pages

893-900

issue

3

eissn

0032-0889

issn

1532-2548

journal_volume

112

pub_type

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